(4-((3r,4r)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2r,6s)-6-(p-tolyl)tetrahydro-2h-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate

ABSTRACT

The invention provides a salt of a tetrahydropyranylmethylaminopyrimidine amide, such as the citrate salt of (4-((3R,4R)-3-methoxytetrahydropyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone, pharmaceutical compositions containing the same, processes for preparing the same, and methods of medical treatment using the same.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/224,902, filed Dec. 19, 2018, which is a continuation of U.S. patentapplication Ser. No. 15/849,929, filed Dec. 21, 2017, now U.S. Pat. No.10,213,428, which is a continuation of International Patent ApplicationNo. PCT/US2016/040728, filed Jul. 1, 2016, which claims the benefit ofand priority to European Patent Application serial number 15175066.8,filed Jul. 2, 2015, the contents of each of which are herebyincorporated by reference in their entirety.

FIELD OF THE INVENTION

The present invention provides the citrate salt of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanoneand to a process for manufacturing it. The present invention alsoprovides the same citrate salt for use in the treatment of medicalconditions, such as acute and chronic mild to moderate musculoskeletalpain, low back pain, chronic low back pain, pain related to rheumatoidarthritis, shoulder pain, dental pain, signs and symptoms ofosteoarthritis, osteoarthritis of the knee, osteoarthritis of the hip,osteoarthritis of the hand, pain associated with osteoarthritis, cancerpain, diabetic polyneuropathy, visceral pain, acute pain, diabeticnephropathy, neuropathic pain, as well as to a pharmaceuticalcomposition comprising the same salt.

BACKGROUND

WO 2011/073154 discloses a number oftetrahydropyranyl-methyl-amino-(hetero)aryl-amides without disclosingany specific salt or crystal form of the compounds exemplified therein.Among others, WO 2011/073154 discloses compound I

Compounds disclosed in WO 2011/073154 are potent CCR2 antagonists.However, in order to prove to be developable for use as a medicament ina human, a drug substance and its solid form must, in addition to invitro and in vivo pharmacokinetic and pharmacological properties andsafety profile, fulfil a series of criteria with regard to therequirements of chemistry, manufacturing and controls (CMC) such assolid form characteristics, purity, drying times, filterability,stability, thermal stability, hygroscopicity, reproducibility andfurther physicochemical properties including solubility and intrinsicdissolution rate.

One of the biggest challenges in the course of the development of a drugproduct for medical use in humans is to identify a drug substance whichis potent, efficacious, fulfils safety requirements and simultaneouslyhas a solid form suitable for human drug development, i.e., fulfillingall the above mentioned criteria cumulatively. This is because each andevery solid form, salt and polymorphic form thereof has physicochemicaland pharmacokinetic properties which are just as unforeseeable asunexpected.

Furthermore, due to the unpredictable and unexpected nature of thesolid, salt and polymorphic forms, there is neither generic nor specificguidance for the skilled person how to design a solid form with thedesired characteristics. Therefore, extensive and creative research andexperimentation is essential to arrive at the specific solid form of aselected drug substance that fulfils all requirements. Optimization ofone crucial parameter often results in the deterioration of another orother parameter(s).

SUMMARY

The objective technical problem underlying the present invention is toprovide a drug substance with CCR2 antagonistic activity which isdevelopable for use as a medicament in humans, i.e., where:

-   a) the drug substance is characterised by high pharmacological    potency, efficacy, in vitro and in vivo pharmacokinetics, and    necessary safety properties; and-   b) the drug substance and its solid form fulfil a series of criteria    with regard to the requirements of chemistry, manufacturing and    controls (CMC) such as solid form characteristics, purity, drying    times, filterability, stability, thermal stability, hygroscopicity,    reproducibility and further physicochemical properties including    solubility and intrinsic dissolution rate.

Compound I has surprisingly been found to fulfil the majority of theabove mentioned criteria required for use as a medicament in humans asdemonstrated (see biological data below). These parameters includeplasma protein binding (relevant for pharmacokinetics andpharmacodynamics), in vitro metabolic stability (relevant forpharmacokinetics), pharmacokinetics and safety properties (hERG,relevant for cardiovascular safety, and drug-induced phospholipidosis).

However, the free base of compound I however turned out to be anamorphous material which was in a metastable state and thus subject tometamorphosis. It was not suitable as a drug substance for developmentbecause it did not fulfil the requirement of being able to bereproducibly manufactured.

Attempts to obtain compound I in crystalline form from solutions in allcommonly used solvents such as ethanol, ethanol/water, 2-propanol,2-propanol/water, acetone, ethyl acetate, 2-butanone or tetrahydrofuranfailed. Such attempts to obtain compound I in crystalline form fromsolutions in all commonly used solvents such as ethanol, ethanol/water,2-propanol, 2-propanol/water, acetone, ethyl acetate, 2-butanone ortetrahydrofuran yielded only amorphous material. Due to these failures,salt forms of compound I with various acids were investigated.

To ensure reproducibility of the physicochemical properties in thepharmaceutical manufacturing process, the drug substance must invariablybe obtained in a well-defined crystalline modification. When acrystalline form of a drug substance or its salt exists in differentpolymorphic modifications (polymorphism), spontaneous conversion of onepolymorphic form into another one may occur. Such a spontaneousinterconversion cannot be tolerated and should be avoided by all means.Therefore, it is essential for securing the reproducibility of thepharmaceutical manufacturing process to identify a salt of a drugsubstance that exists either in one crystalline form only, or that atleast is characterized by a reduced tendency towards polymorphism.

According to the present invention, the technical problem outlined abovehas been solved by experimentation and innovation that resulted in theidentification of the specific compound(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate salt 1

The citrate salt 1 is crystalline, i.e., defined by a specific crystalmodification, thus allowing to obtain the drug substance in high purityand reproducibly high stability.

Various salt forms of compound I where prepared and analysed. Forinstance, crystalline forms of the citrate, hydrobromide, hydrochloride,esilate and methanesulfonate salt of compound I were obtained bycrystallization. Analysis of these salt forms unexpectedly revealed thatthe citrate, esilate and methanesulfonate salts of compound I exhibitedonly one polymorphic form. This stands in contrast to the hydrobromideand hydrochloride salts of compound I, which were obtained in differentpolymorphic modifications.

Another key parameter of a drug substance is hygroscopicity. Wateruptake of a salt of a drug substance by sorption during manufactureleads to a reduced amount of the drug substance in the drug product andtherefore to reduced efficacy. In addition, water uptake of a salt of adrug substance or a drug product may lead to decomposition of the drugsubstance. Therefore, it is essential to identify a drug substance orsalt thereof that is not hydroscopic, or has only very littlehygroscopic character.

Unexpectedly, the crystalline form of the citrate salt 1 of compound Iis characterized by low and reversible water uptake at a relativehumidity of up to 90% (2.6% water uptake at 80% relative humidity and3.4% water uptake at 90% relative humidity). On the contrary, thecrystalline forms of the corresponding hydrobromide, hydrochloride,esilate and methanesulfonate of compound I readily absorb significantamounts of water at a relative humidity of as low as 80% and becomeirreversibly deliquescent.

Accordingly, one aspect of the invention provides the compound havingthe following formula 1:

In certain embodiments, the compound is provided in crystalline form.

Another aspect of the invention provides a pharmaceutical compositioncomprising (i) a compound described herein such as citrate salt 1 and(ii) one or more carriers and/or diluents. In certain embodiments, thepharmaceutical composition is formulated for oral administration.

Another aspect of the invention provides the citrate salt 1 or apharmaceutical composition comprising said citrate salt 1 for use intreating a medical condition. Exemplary medical conditions include, forexample, treatment of pain (e.g., inflammatory pain or neuropathic pain)and osteoarthritis.

Another aspect of the invention provides a method of treating a medicalcondition in a patient, where the method comprises administering to apatient in need thereof a therapeutically effective amount of a compounddescribed herein, such as citrate salt 1, in order to treat the medicalcondition. Exemplary medical conditions include, for example, pain(e.g., inflammatory pain or neuropathic pain) and osteoarthritis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the X-ray powder diffractogram of the amorphous base of thecompound(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)pipendin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.

FIG. 2 shows the X-ray powder diffractogram of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-yl)amino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonecitrate.

FIG. 3 shows the thermoanalysis and determination of the melting point(DSC/TG) of (4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate.

FIG. 4 shows the sorption isotherms of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate.

FIG. 5 shows the X-ray powder diffractogram of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonehydrobromide.

FIG. 6 shows the thermoanalysis and determination of the melting point(DSC/TG) of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrobromide.

FIG. 7 shows the sorption isotherms of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrobromide.

FIG. 8 shows the X-ray powder diffractogram of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonehydrochloride.

FIG. 9 shows the thermoanalysis and determination of the melting point(DSC/TG) of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrochloride.

FIG. 10 shows the sorption isotherms of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrochloride.

FIG. 11 shows the X-ray powder diffractogram of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-yl)amino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanoneesilate.

FIG. 12 shows the thermoanalysis and determination of the melting point(DSC/TG) of (4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone esilate.

FIG. 13 shows the sorption isotherms of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone esilate.

FIG. 14 shows the X-ray powder diffractogram of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonemethanesulfonate.

FIG. 15 shows the thermoanalysis and determination of the melting point(DSC/TG) of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-l-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonemethanesulfonate.

FIG. 16 shows the sorption isotherms of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone methanesulfonate.

FIG. 17 shows the FT-RAMAN spectrum of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate.

FIG. 18 shows the FT-RAMAN spectrum of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone esilate.

DETAILED DESCRIPTION

The invention provides salt forms of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone,pharmaceutical compositions containing such salt forms, methods forpreparing salt forms, and therapeutic methods for using such salt forms,such as in the treatment of pain and other medical conditions. Asdescribed herein, the citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone was surprisingly discovered to provide multipleunexpected benefits over other salt forms of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.For example, the citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone was found to exhibit low andreversible water uptake at a relative humidity up to 90%, which standsin contrast to salt forms of the corresponding hydrobromide,hydrochloride, esilate, and methanesulfonate that readily absorbsignificant amounts of water at a relative humidity of as low as 80% andbecome irreversibly deliquescent. Further still, the citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonewas unexpectedly found to exhibit only one polymorphic crystalline form,which stands in contrast to the corresponding crystalline salts formedfrom hydrobromic acid and hydrochloric acid that exhibited differentpolymorphic modifications. Accordingly, the citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanoneis surprisingly superior for development as a pharmaceutical due to themultiple unexpected properties that are beneficial.

Various aspects and embodiments of the invention are further describedbelow in sections. Aspects and embodiments of the invention described inone particular section are not to be limited to any particular section.

To facilitate an understanding of the present invention, a number ofterms and phrases are defined below.

The terms “subject” and “patient” refer to organisms to be treated bythe methods of the present invention. Such organisms include mammals(e.g., murines, simians, equines, bovines, porcines, canines, felines,and the like), and most preferably is humans.

The term “effective amount” refers to the amount of a compoundsufficient to effect beneficial or desired results. An effective amountcan be administered in one or more administrations, applications ordosages and is not intended to be limited to a particular formulation oradministration route.

The term “treating” includes any effect, e.g., lessening, reducing,modulating, ameliorating or eliminating, that results in the improvementof the condition, disease, disorder, and the like, or ameliorating asymptom thereof.

Throughout the description, where compositions are described as having,including, or comprising specific components, or where processes andmethods are described as having, including, or comprising specificsteps, it is contemplated that, additionally, there are compositions ofthe present invention that consist essentially of, or consist of, therecited components, and that there are processes and methods accordingto the present invention that consist essentially of, or consist of, therecited processing steps.

I. Salt Forms of4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone

One aspect of the invention provides salt forms of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.As described below and in the working examples, this disclosuredescribes salt forms of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanoneprepared by reacting4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone with an acid selected fromcitric acid, hydrobromic acid, hydrochloric acid, ethanesulfonic acid,and methanesulfonic acid.

The citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone was surprisingly discovered toprovide multiple unexpected benefits over other salt forms of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone. For example, the citrate saltof4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone was found to exhibit low andreversible water uptake at a relative humidity up to 90%, which standsin contrast to salt forms of the corresponding hydrobromide,hydrochloride, esilate, and methanesulfonate that readily absorbsignificant amounts of water at a relative humidity of as low as 80% andbecome irreversibly deliquescent. Further still, the citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonewas unexpectedly found to exhibit only one polymorphic crystalline form,which stands in contrast to the corresponding crystalline salts formedfrom hydrobromic acid and hydrochloric acid that exhibited differentpolymorphic modifications. Accordingly, the citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanoneis surprisingly superior for development as a pharmaceutical due to themultiple unexpected properties that are beneficial.

Accordingly, one aspect of the invention provides the citrate salt ofcompound I:

having the formula

In certain embodiments, said citrate salt is in crystalline form.

In certain embodiments, the crystalline form is characterized by showinga X-ray powder diffraction pattern comprising peaks at the following2-theta values measured using monochromatic CuKα1 radiation of λ=1.54056Å, 40 kV, 40 mA: 19.1° and 22.4°. In certain embodiments, thecrystalline form is characterized in that the X-ray powder diffractionpattern further comprises a peak at 12.2°. In certain embodiments, thecrystalline form is, characterized in that the X-ray powder diffractionpattern further comprises a peak at 13.7°. In certain embodiments, thecrystalline form is characterized in that the X-ray powder diffractionpattern further comprises a peak at 14.6°. In certain embodiments, thecrystalline form is characterized in that the X-ray powder diffractionpattern further comprises a peak at 18.7°. In certain embodiments, thecrystalline form is characterized in that the X-ray powder diffractionpattern further comprises a peak at 24.6°. In certain embodiments, thecrystalline form is characterized in that X-ray powder diffractionpattern further comprises a peak at 26.3°.

In certain embodiments, the crystalline form exhibits a X-ray powderdiffraction pattern comprising peaks at the following 2-theta valuesmeasured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40 kV, 40mA: 12.2±0.2, 13.7±0.2, 14.6±0.2, 19.1±0.2, and 22.4±0.2. In certainother embodiments, the crystalline form exhibits a X-ray powderdiffraction pattern comprising peaks at the following 2-theta valuesmeasured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40 kV, 40mA: 12.2±0.2, 13.7±0.2, 14.6±0.2, 18.7±0.2, 19.1±0.2, 22.4±0.2,24.6±0.2, and 26.3±0.2.

In certain embodiments, the relative intensity of the peak at saiddiffraction angles 2-theta is at least 10%. In certain otherembodiments, the relative intensity of the peak at said diffractionangles 2-theta is at least 15%.

In certain embodiments, the crystalline form has a X-ray powderdiffraction pattern that is substantially as shown in FIG. 2.

In certain embodiments, the crystalline form is characterized by thefollowing X-ray powder diffraction pattern expressed in terms ofdiffraction angle 2θ, inter-planar distances d, and relative intensity(expressed as a percentage with respect to the most intense peak):

2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.36 20.24 17 12.17 7.27 4112.51 7.07 6 13.13 6.74 7 13.66 6.48 39 14.20 6.23 14 14.60 6.06 3215.03 5.89 5 15.25 5.81 4 15.97 5.54 11 16.51 5.37 13 17.05 5.20 1317.54 5.05 4 17.88 4.96 5 18.65 4.75 22 19.05 4.66 100 19.68 4.51 1120.42 4.35 6 20.84 4.26 4 21.25 4.18 3 21.90 4.06 5 22.42 3.96 92 23.193.83 9 23.70 3.75 16 24.34 3.65 4 24.56 3.62 23 24.89 3.57 16 25.20 3.537 25.36 3.51 7 25.67 3.47 6 26.26 3.39 23 26.59 3.35 12 27.51 3.24 627.71 3.22 6 28.01 3.18 7 28.23 3.16 5 28.57 3.12 3 29.44 3.03 12 30.152.96 4

The crystalline form may be further characterized according to its Ramanspectrum. Accordingly, in certain embodiments, the crystalline form hasa Raman spectrum comprising peaks at any one or all of the followingRaman shifts expressed in wavenumbers in cm⁻¹: 1718, 1242, 731, 662,553.

The crystalline form may be further characterized according to itsmelting point. Accordingly, in certain embodiments, the crystalline formhas a melting point of 212±5° C.

The crystalline form may be further characterized according to itsdifferential scanning calorimetry curve. Accordingly, in certainembodiments, the crystalline form has a differential scanningcalorimetry curve substantially the same as shown in FIG. 3.

Desirably the molar ratio of citric acid to4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone is about 1:1 in acitrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.In certain embodiments, the molar ratio of citric acid to4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone is in the range of 1.2:1to 1:1.2 in a citrate salt of4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone. In certain other embodiments, the molar ratio of citricacid to4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone is 1:1 in a citrate saltof4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.

The compound(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone (I) is specifically disclosed inWO 2011/073154, as well as a process for its preparation. For details ona process to manufacture this compound, reference is thus made to WO2011/073154 (example 30, page 150).

Methods for preparing citrate salt 1 are also provided. For example, oneaspect of the invention provides a method for preparing compound 1

comprising the following steps:

-   -   a) addition of citric acid to a solution of compound I

-   -   in an organic solvent    -   b) isolation of the resulting salt 1 in pure form.

In certain embodiments, the method is further characterized in that theorganic solvent in step a) is selected from the group consisting ofethyl acetate, isopropanol and a mixture of isopropanol and water.

II. Therapeutic Applications

Compounds such as those described in Section I (e.g., citrate salt 1)and pharmaceutical compositions described herein are useful as amedicament. The medicament may be for treating a disorder in whichinhibition of CCR2 activity provides a therapeutic benefit.

The Chemokine receptor CCR2 has been reported to be implicated as beingan important mediator of inflammatory and immunoregulatory disorders anddiseases as well as autoimmune pathologies such as rheumatoid arthritisand atherosclerosis. See, for example, WO 2010/070032. Thus, agents thatmodulate the chemokine receptor CCR2 are useful in treating suchdisorders and diseases.

More generally, it is widely accepted that numerous conditions anddiseases involve inflammatory processes. Such inflammations arecritically triggered and/or promoted by the activity of macrophages,which are formed by differentiation out of monocytes. It has furtherbeen found that monocytes are characterized by, e.g., a high expressionof membrane-resident CCR2, whereas the CCR2 expression in macrophages islower. CCR2 is a critical regulator of monocytes trafficking, which canbe described as the movement of the monocytes towards an inflammationalong a gradient of monocyte chemoattractant proteins (MCP-1, MCP-2,MCP-3, MCP-4).

Therefore, in order to reduce macrophage-induced inflammation, it wouldbe desirable to block the monocyte CCR2 by an antagonist, so that themonocytes can be less triggered to move towards an inflammation area forconversion into macrophages.

Accordingly, one aspect of the invention provides a method of treating aCCR2-related condition in a patient, where the method comprisesadministering to a patient in need thereof a therapeutically effectiveamount of a compound described herein (e.g., the citrate salt 1 or acrystalline form thereof) to treat the condition. In certainembodiments, the CCR2-related condition is a MCP-1 related condition.

In certain embodiments, the CCR2-related condition is pain. Exemplarytypes of pain contemplated for treatment, include, for example,inflammatory pain, neuropathic pain, and visceral pain. In certainembodiments, the pain is chronic pain. In certain embodiments, the painis pain due to osteoarthritis. Other exemplary types of paincontemplated for treatment include, for example, low back pain, hippain, leg pain, non-herpetic neuralgia, post herpetic neuralgia,diabetic neuropathy, nerve injury-induced pain, acquired immunedeficiency syndrome (AIDS) related neuropathic pain, head trauma, toxinand chemotherapy caused nerve injuries, phantom limb pain, painfultraumatic mononeuropathy, painful polyneuropathy, thalamic painsyndrome, post-stroke pain, central nervous system injury, post surgicalpain, carpal tunnel syndrome, trigeminal neuralgia, post mastectomysyndrome, postthoracotomy syndrome, stump pain, repetitive motion pain,neuropathic pain associated hyperalgesia and allodynia, alcoholism andother drug-induced pain.

In certain other embodiments, the CCR2-related condition is an immunerelated disease. Exemplary immune-related diseases include, for example,rheumatoid arthritis, juvenile rheumatoid arthritis, systemic onsetjuvenile rheumatoid arthritis, psoriatic arthritis, ankylosingspondilitis, gastric ulcer, seronegative arthropathies, osteoarthritis,inflammatory bowel disease, and ulcerative colitis.

In certain other embodiments, the CCR2-related condition is a fibroticcondition. Exemplary fibrotic conditions include, for example, liverfibrosis (including but not limited to alcohol-induced cirrhosis,viral-induced cirrhosis, autoimmune-induced hepatitis); lung fibrosis(including but not limited to scleroderma, idiopathic pulmonaryfibrosis); kidney fibrosis (including but not limited to scleroderma,diabetic nephritis, glomerular pehpritis, lupus nephritis); dermalfibrosis (including but not limited to scleroderma, hypertrophic andkeloid scarring, burns); myelofibrosis; neurofibromatosis; fibroma;intestinal fibrosis; and fibrotic adhesions resulting from surgicalprocedures.

In certain other embodiments, the CCR2-related condition is aninflammatory disorder.

Another aspect of the invention provides a method of treating acondition selected from pain, osteroarthritis, diabetic nephropathy, anddiabetic polyneuropathy, where the method comprises administering to apatient in need thereof a therapeutically effective amount of a compounddescribed herein (e.g., the citrate salt 1 or a crystalline formthereof) to treat the condition.

In certain embodiments, the condition is pain. In certain embodiments,the condition is inflammatory pain. In certain embodiments, thecondition is chronic pain. In certain embodiments, the condition is paindue to osteoarthritis. In certain embodiments, the condition isneuropathic pain or visceral pain.

In certain embodiments, the condition is selected from the groupconsisting of acute and chronic mild to moderate musculoskeletal pain,low back pain, chronic low back pain, pain related to rheumatoidarthritis, shoulder pain, dental pain, signs and symptoms ofosteoarthritis, osteoarthritis of the knee, osteoarthritis of the hip,osteoarthritis of the hand, pain associated with osteoarthritis, cancerpain, diabetic polyneuropathy, visceral pain, acute pain, diabeticnephropathy, and neuropathic pain. In certain embodiments, the conditionis pain selected from (a) trigeminal neuralgia and (b) pain due tochemotherapy caused nerve injury.

In certain embodiments, the condition is osteoarthritis.

In certain embodiments, the method comprises administering to thepatient a therapeutically effective amount of citrate salt 1 to treatthe condition.

In a more specific embodiment, the invention provides for using acompound described herein for the treatment of a disease in whichinhibition of the CCR2 receptor is beneficial, such as: (i) acute andchronic mild to moderate musculoskeletal pain (low back pain, chroniclow back pain, pain related to rheumatoid arthritis, shoulder pain,dental pain); (ii) signs and symptoms of osteoarthritis (osteoarthritisof the knee and/or hip, osteoarthritis of the hand, pain associated withosteoarthritis); (iii) cancer pain; (iv) diabetic polyneuropathy; (v)visceral pain, (vi) acute pain, (vii) diabetic nephropathy; and (viii)neuropathic pain.

III. Pharmaceutical Compositions

Another aspect of the invention provides a pharmaceutical compositioncomprising a compound described herein (e.g., citrate salt 1) togetherwith one or more inert carriers and/or diluents. The pharmaceuticalcompositions may be formulated for administration via a particularroute, such as oral administration.

More generally, suitable forms for administration are, for example,tablets, capsules, solutions, syrups, emulsions or inhalable powders oraerosols. The content of the pharmaceutically effective compound(s) ineach case should be in the range from 0.1 to 90 wt. %, preferably 0.5 to50 wt. % of the total composition, i.e., in amounts which are sufficientto achieve the dosage range specified hereinafter.

The preparations may be administered orally in the form of a tablet, asa powder, as a powder in a capsule (e.g. a hard gelatine capsule), as asolution or suspension. When administered by inhalation the activesubstance combination may be given as a powder, as an aqueous oraqueous-ethanolic solution or using a propellant gas formulation.

Suitable tablets may be obtained, for example, by mixing the activesubstance(s) with known excipients, for example inert diluents such ascalcium carbonate, calcium phosphate or lactose, disintegrants such ascorn starch or alginic acid, binders such as starch or gelatine,lubricants such as magnesium stearate or talc and/or agents for delayingrelease, such as carboxymethyl cellulose, cellulose acetate phthalate,or polyvinyl acetate. The tablets may also comprise several layers.Coated tablets may be prepared accordingly by coating cores producedanalogously to the tablets with substances normally used for tabletcoatings, for example collidone or shellac, gum arabic, talc, titaniumdioxide or sugar. To achieve delayed release or preventincompatibilities the core may also consist of a number of layers.Similarly the tablet coating may consist of a number of layers toachieve delayed release, possibly using the excipients mentioned abovefor the tablets.

Syrups containing the active substances or combinations thereofaccording to the invention may additionally contain a sweetener such assaccharine, cyclamate, glycerol or sugar and a flavour enhancer, e.g. aflavouring such as vanillin or orange extract. They may also containsuspension adjuvants or thickeners such as sodium carboxymethylcellulose, wetting agents such as, for example, condensation products offatty alcohols with ethylene oxide, or preservatives such asp-hydroxybenzoates.

Capsules containing one or more active substances or combinations ofactive substances may for example be prepared by mixing the activesubstances with inert carriers such as lactose or sorbitol and packingthem into gelatine capsules.

Suitable suppositories may be made for example by mixing with carriersprovided for this purpose, such as neutral fats or polyethyleneglycol orthe derivatives thereof

Excipients which may be used include, for example, water,pharmaceutically acceptable organic solvents such as paraffins (e.g.petroleum fractions), vegetable oils (e.g. groundnut or sesame oil),mono- or polyfunctional alcohols (e.g. ethanol or glycerol), carrierssuch as e.g. natural mineral powders (e.g. kaolins, clays, talc, chalk),synthetic mineral powders (e.g. highly dispersed silicic acid andsilicates), sugars (e.g. cane sugar, lactose and glucose), emulsifiers(e.g. lignin, spent sulphite liquors, methylcellulose, starch andpolyvinylpyrrolidone) and lubricants (e.g. magnesium stearate, talc,stearic acid and sodium lauryl sulphate).

For oral administration, the tablets may, of course, contain, apart fromthe abovementioned carriers, additives such as sodium citrate, calciumcarbonate and dicalcium phosphate together with various additives suchas starch, preferably potato starch, gelatine and the like. Moreover,lubricants such as magnesium stearate, sodium lauryl sulphate and talcmay be used at the same time for the tabletting process. In the case ofaqueous suspensions the active substances may be combined with variousflavour enhancers or colourings in addition to the excipients mentionedabove.

IV. Kits for Use in Medical Applications

Another aspect of the invention provides a kit for treating a medicalcondition. The kit comprises: i) instructions for treating a medicalcondition, such as pain, osteroarthritis, diabetic nephropathy, ordiabetic polyneuropathy (for example, pain such as selected from acuteand chronic mild to moderate musculoskeletal pain, low back pain,chronic low back pain, pain related to rheumatoid arthritis, shoulderpain, dental pain, pain associated with osteoarthritis, cancer pain,visceral pain, acute pain, diabetic nephropathy, and neuropathic pain);and ii) a compound described herein, such as citrate salt 1. The kit maycomprise one or more unit dosage forms containing an amount of citratesalt 1 that is effective for treating said medical condition, such aspain.

EXAMPLES

The invention now being generally described, will be more readilyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention.

List of Abbreviations

-   AUC area under the plasma concentration-time curve-   BR hydrobromide (salt with hydrobromic acid)-   BS base (no salt defined)-   C_(max) peak concentration-   CI citrate (salt with citric acid)-   CL clearance-   CL hydrochloride (salt with hydrochloric acid)-   ES esilate (salt with one mol ethanesulfonic acid)-   d.b. (on) dry basis-   DSC Differential Scanning calorimeter-   DMSO dimethyl sulfoxide-   DMSO-d₆ deuterated DMSO-   DVS Dynamic vapour sorption-   EDTA Ethylenediaminetetraacetic acid-   EGTA ethylene glycol tetraacetic acid-   ESI electrospray ionization-   f female-   F oral bioavailability-   FCS fetal calf serum-   HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid-   hERG human ether-a-go-go-related gene-   HR high-resolution-   IV, i.v. intravenous-   m male-   M mol/L-   Mcllvaine buffer citrate/phosphate buffer-   MRT_(disp) mean residence time following intravenous dosing-   MRT_(tot) mean residence time following oral dosing-   MS mass spectrometry-   MS methanesulfonate (salt with one mol methanesulfonic acid)-   m/z mass-to-charge ratio-   NMR nuclear magnetic resonance-   PBS phosphate buffered saline-   PK pharmacokinetics-   PO, p.o. peroral-   r.h. relative humidity-   RT room temperature-   Sorensen buffer NaOH/NaCL/Glycin-buffer-   t_(max) time of maximum plasma concentration-   TG ThermoGravimetry-   V_(ss) steady-state volume of distribution-   VLE very low endotoxin-   XRPD X-ray powder diffraction

EXAMPLE 1—Preparation and Physicochemical Characterization of Salts ofCompound I

Multiple salts of compound I were prepared and characterized, includingthe citrate salt of compound I. Experimental procedures and results areprovided below. Compound I has the following formula:

Part I: Description of Analytical Methods Used

Provided below is a description of analytical methods used tocharacterize salts of compound I.

ESI Mass Spectrometry (ER+)

Instrument QTOF 2 (Micromass, Manchester, UK) Instrument controlsoftware Masslynx 4.1 Ion source ESI + (Lockspray source) Lockspray/DXCon/off Calibration 0.1% Phosphoric acid in acetonitrile/ water (1:1),lockmass calibration Resolution MS1(LM/HM) 5/5 Resolving power (FWHM)16000 at m/z 491 (W mode) MCP voltage 2200 V Capillary voltage +2.8 kVCone voltage 25 V Collision energy 5 V Collision gas Argon Sourcetemperature 120° C. Desolvation temperature 150° C. Cone gas nitrogen 75L/h Desolvation gas nitrogen 450 L/h Spray solvent acetonitrile/water9:1 Syringe pump Harvard Apparatus 55-2222 Spray solvent flow rate 5μL/min Sample concentration 5 ng/μL spray solvent Reagents acetonitrile(ULC/MS, Biosolve) water (purified by Milli-Q-system) Scan range 50-1000u (TOF scan, profile data) Scan time 2.9 s No. of scans combined 20Accurate mass determination Center 5 points/80%, Np = 0.35, lockmass:588.8692 Data threshold 1.0%

¹NMR Spectroscopy

Instrument Bruker DRX 400 Frequency 400.13 MHz Software TopSpin ®version 1.3 PL8 Pulse program zg30 Solvent DMSO-d₆ Concentration 10.3mg/0.6 mL Temperature 30° C. Calibration TMS (δ = 0.00 ppm) Sweep width8013 Hz Size 64K data points Pulse width 30 degree Relaxation delay 10 sNumber of scans 32 Dummy scans 8 Apodization zerofilling to 128K datapoints Gaussian multiplication (GB: 0.25, LB: −0.25 Hz)

¹³C NMR Spectroscopy

Instrument Bruker DRX 400 Frequency 100.61 MHz Software TopSpin ®version 1.3 PL8 Pulse program Zgpg Solvent DMSO-d₆ Concentration 10.3mg/0.6 ml Temperature 30° C. Calibration DMSO-d₆ (δ = 39.5 ppm) Sweepwidth 27778 Hz Size 64K data points Pulse width 90 degree Relaxationdelay 4 s Number of scans 4096 Dummy scans 32 Apodization zerofilling to128K data points Exponential multiplication (LB: 2.5 Hz)

X-Ray Powder (XRPD) Diagram

X-ray powder diagrams were generated using a STOE—STADI P-diffractometerin transmission mode fitted with a MYTHEN-detector and a Cu-anode asX-ray source with monochromatic CuKα1 radiation (λ=1.54056 Å, 40 kV, 40mA).

FT-RAMAN Spectroscopy

Samples have been measured in boiling point tubes using a Bruker RAM IIFT-Raman Module instrument, resolution 2 cm⁻¹, 64 scans, laser power 500mW (focussed laser). Analysis: scaling of vector in spectral range 3500cm⁻¹-50 cm⁻¹.

Differential Scanning Calorimetry—Melting Point

The compounds are characterised by a melting point determined byDifferential Scanning calorimetry (DSC), evaluated by the peak maximumor onset temperature. The heating rate of the experiment is 10° C./min.The values given were determined using a DSC instrument from theQ-series™ of TA Instruments.

ThermoGravimetry (TG)

Thermal gravimetry data were collected with a TG instrument from theQ-series of TA Instruments. This method measures weight changes in amaterial as a function of temperature under a controlled atmosphere.

Dynamic Vapour Sorption (DVS)

Sorption isotherms were generated using an IGAsorp water sorptionmonitor from Hiden Isochema. Adsorption and desorption isotherms wereobtained at 25° C. with 10% r.h. step intervals ranging from 10% r.h. to90% r.h.

For BR salt form only: Sorption isotherms were registered on a DVS-1water sorption monitor from Surface Measurement Systems.

Solubility

Solubility was determined using an automated shake flask method (at roomtemperature) and quantitation of the dissolved drug substance wasdetermined by UV-spectroscopy within this automated setup.

Part II: Preparation of(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate (1)

Exemplary procedures for making the title compound are provided below,along with physical characterization data. The preparation proceduresinclude two different routes for making the title compound.

Preparation Option a) Preparation of Citrate Salt Starting from FreeBase I

To a solution of the free base I (200 mg, 0.372 mmol) in ethyl acetate(2 mL) is added citric acid mono hydrate (78.2 mg; 0.372 mmol). Thesolution is stirred overnight (18 h). The suspension is filtered and theproduct is dried at 40° C. in vacuo to yield 140 mg 0.192 mmol (52%)colourless crystals. Physical characterization data for citrate salt 1is provided below.

NMR (¹H, 400 MHz, DMSO-d₆): 11.7-8.5; (2H, broad), 8.34; (1H, s), 7.22;(2H, m), 7.12; (2H, m), 7.08; (1H, t), 4.49; (1H, m), 4.31; (1H, d),4.09; (1H, m), 3.85; (1H, m), 3.74; (1H, m), 3.57-3.44; (2H, m), 3.48;(1H, m), 3.47; (1H, m), 3.35; (3H, s), 3.35; (1H, m), 3.33 (1H, m),3.29; (1H, m), 3.27; (1H, m), 3.04; (1H, m), 2.84; (1H, m), 2.58; (2H,d), 2.50; (2H, d), 2.28; (3H, s), 2.12; (1H, m), 1.94; (1H, m), 1.91;(3H, s), 1.88; (1H, m), 1.78; (1H, m), 1.76 (1H, m), 1.70; (1H, m),1.66; (1H, m), 1.63; (1H, m), 1.40; (1H, m), 1.40; (1H, m), 1.37; (1H,m), 1.24; (1H, m) (includes rotamers).

NMR (¹³C, 100 MHz, DMSO-d₆): 176.6, 171, 165.4, 161.0, 156.6, 155.4,140.3, 136.0, 128.5, 125.6, 109.3, 78.5, 75.4, 72.4, 72.2, 71.2, 64.8,64.4, 64.4, 55.5, 55.5, 51.5, 51.4, 50.2, 45.6, 44.1, 44.1, 38.8, 33.3,29.6, 28.7, 28.7, 25.1, 23.1, 20.6, 11.7 (includes rotamers).

HRMS (ESI): m/z 538.3400 ([M +H]⁺; C₃₀H₄₄N₅O₄).

FT-RAMAN spectrum (characteristic bands) [cm⁻¹]: 1718, 1242, 731, 662,553.

See table II below and FIGS. 2-4 and 17 for additional characterizingdata.

Preparation Option b) Amide Coupling Followed by Preparation Of CitrateSalt

4.99 kg (30.75 mol) of 1,1′-carbonyldiimidazole are added to asuspension of 10.0 kg (29.29 mol) of 2 in 75 L of2-methyltetrahydrofuran at 50° C. The powder funnel is rinsed with 5 L2-methyltetrahydrofuran. The reaction mixture is stirred for 70 min at50° C. Then, 8.83 kg (30.75 mol) of 3 are added to the reaction mixtureand the funnel is rinsed with 5 L 2-methyltetrahydrofuran. Next, 7.41 kg(73.23 mol) of triethylamine and 10 L of 2-methyltetrahydrofuran areadded and the reaction mixture is stirred for 1 h under reflux. Then,the mixture is cooled to 60° C. and a solution of 6.07 kg (43.94 mol) ofpotassium carbonate in 55 L water is added and the phases are separatedat 55° C. The organic layer is washed with 60 L water and 80 L ofsolvent are removed by distillation in vacuo. The resulting residue isdiluted with 80 L of isopropyl alcohol and 55 L of solvent is removed bydistillation in vacuo. The resulting residue is diluted with 40 L ofisopropyl alcohol and 40 L of solvent is removed by distillation invacuo. Next, 5.85 kg (27.83 mol) of citric acid monohydrate in 11 L ofwater are added and the dropping funnel is rinsed with 30 L of isopropylalcohol. The reaction mixture is heated to 75° C., stirred until asolution is formed, and then filtrated. The filter is rinsed with amixture of 2 L of water and 20 L of isopropyl alcohol. Then, thefiltrate is diluted with 30 L of isopropyl alcohol and seeded with 100 gof 1 as obtained in option a) at 65° C. Next, the mixture is cooled to55° C. within 30 minutes and then further stirred for 1 h at 55° C. Theresulting suspension is diluted with 60 L of isopropyl alcohol within 1h at 55° C. and then cooled to 20° C. within 3 h. Then, the suspensionis stirred for 17 h at 20° C. and isolated by filtration. The filtercake is washed twice with a mixture of 19 L of isopropyl alcohol and 1 Lof water, each. The product is dried at 50° C. in vacuo to yield 17.76kg of compound (83%). Physical characterization data for citrate salt 1is provided below.

NMR (¹H, 400 MHz, DMSO-d₆): 11.7-8.5; (2H, broad), 8.34; (1H, s), 7.22;(2H, m), 7.12; (2H, m), 7.08; (1H, t), 4.49; (1H, m), 4.31; (1H, d),4.09; (1H, m), 3.85; (1H, m), 3.74; (1H, m), 3.57-3.44; (2H, m), 3.48;(1H, m), 3.47; (1H, m), 3.35 (3H, s), 3.35; (1H, m), 3.33 (1H, m), 3.29;(1H, m), 3.27; (1H, m), 3.04; (1H, m), 2.84; (1H, m), 2.58; (2H, d),2.50; (2H, d), 2.28; (3H, s), 2.12; (1H, m), 1.94; (1H, m), 1.91; (3H,s), 1.88; (1H, m), 1.78; (1H, m), 1.76; (1H, m), 1.70; (1H, m), 1.66(1H, m), 1.63; (1H, m), 1.40; (1H, m), 1.40; (1H, m), 1.37; (1H, m),1.24; (1H, m) (includes rotamers).

NMR (¹³C, 100 MHz, DMSO-d₆): 176.6, 171, 165.4, 161.0, 156.6, 155.4,140.3, 136.0, 128.5, 125.6, 109.3, 78.5, 75.4, 72.4, 72.2, 71.2, 64.8,64.4, 64.4, 55.5, 55.5, 51.5, 51.4, 50.2, 45.6, 44.1, 44.1, 38.8, 33.3,29.6, 28.7, 28.7, 25.1, 23.1, 20.6, 11.7 (includes rotamers).

HRMS (ESI): m/z 538.3400 ([M +H]⁺; C₃₀H₄₄N₅O₄).

FT-RAMAN spectrum (characteristic bands) [cm⁻¹]: 1718, 1242, 731, 662,553.

See table II below and FIGS. 2-4 and 17 for additional characterizingdata.

Part III: Preparation of Additional Salts of(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone

Additional salts of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone were prepared and characterizedas described below.

Preparation of(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrobromide

1.916 mL (0.1 M) of hydrobromic acid is added to a solution of 103 mg(0.1916 mmol) of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,65)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone in 2 mL of methanol and stirredfor 2 h at 50° C. Then, the solvent is removed in a vacuum dryer at 40°C. Next, 4 mL of tetrahydrofuran is added to the residue. The mixture issonicated, then stirred for 2 h at 40° C., and afterwards stored for 4 hat room temperature. Then, the solvent is removed in a vacuum dryer toyield the hydrobromide of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.

See table III below and FIGS. 5-7 for characterizing data.

Preparation of(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrochloride

0.558 mL (0.1 M) of hydrochloric acid is added to a solution of 30 mg(0.0557 mmol) of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,65)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone in 1 mL of methanol and stirredfor 2 h at 50° C. Then, the solvent is removed in a vacuum dryer at 40°C. Next, 1.2 mL of tetrahydrofuran is added to the residue. The mixtureis sonicated, then stirred for 2 h at 40° C., and afterwards stored for4 h at room temperature. Then, the solvent is removed in a vacuum dryerto yield the hydrochloride of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,65)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.

See table IV below and FIGS. 8-10 for characterizing data.

Preparation of(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone esilate

1.860 mL (0.1 M) of ethanesulfonic acid is added to a solution of 100 mg(0.186 mmol) of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,65)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone in 2 mL of methanol and stirredfor 2 h at 50° C. Then, the solvent is removed in a vacuum dryer at 40°C. Next, 4 mL of acetone is added to the residue. The mixture issonicated, then stirred for 2 h at 40° C., and afterwards stored for 4 hat room temperature. Then, the solvent is removed in a vacuum dryer toyield the esilate of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.

FT-RAMAN spectrum (characteristic bands) [cm⁻¹]: 1637, 1253, 1014, 740,719, 534, 525, 219.

See table V below and FIGS. 11-13 and 18 for characterizing data.

Preparation of(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone methanesulfonate

0.558 mL (0.1 M) of methanesulfonic acid is added to a solution of 30 mg(0.0557 mmol) of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone in 1 mL methanol and stirred for2 h at 50° C. Then, the solvent is removed in a vacuum dryer at 40° C.Next, 1.2 mL toluene is added to the residue. The mixture is sonicated,then stirred for 2 h at 50° C., and afterwards stored over night at roomtemperature. Then, the solvent is removed in a vacuum dryer to yield themethanesulfonate of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone.

See table VI below and FIGS. 14-16 for characterizing data.

Part IV: Physical Characterization Data for Salts of(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-())2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone

Exemplary physical characterization data for salts of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone is provided below.

Solubility in Aqueous Media

Table I shows the solubility of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonecitrate in different aqueous media at 2, 4 and 6 h.

TABLE I Medium 2 h [mg/ml] 4 h [mg/ml] 6 h [mg/ml] Water >1 >1 >1 0.1NHCl >1 >1 >1 0.01N HCl >1 >1 >1 McIlvaine buffer pH 2.2 >1 >1 >1McIlvaine buffer pH 3.0 >1 >1 >1 McIlvaine buffer pH 4.0 >1 >1 >1McIlvaine buffer pH 4.5 Not determined >1 >1 McIlvaine buffer pH5.0 >1 >1 >1 McIlvaine buffer pH 6.0 >1 >1 >1 McIlvaine buffer pH6.8 >1 >1 >1 McIlvaine buffer pH 7.4 >1 >1 >1 KH₂PO₄-buffer pH7.4 >1 >1 >1 Sörensen pH 10 >1 >1 >1 0.1N NaOH >1 >1 >1 EtOH 9.2 9.8 10

The data in table I demonstrate that(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate is highly soluble inacidic, neutral and basic aqueous media.

Solid State Properties of Citrate Salt 1

Various solid state properties of citrate salt 1 are described below.

Appearance

In the solid state,(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate is a whitemicrocrystalline material.

Sorption Behaviour

Only(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone citrate shows stability against relative humidity up to80%. An uptake of 2.6% water is observed. The water uptake isreversible, and after the sorption experiment the compound still remainsas solid material. All other salts turned into liquid phase at higherrelative humidity (depending on the salt form starting at 60-70%relative humidity).

Crystallinity and Polymorphism Citrate Salt 1

Citrate salt 1 is highly crystalline as can be seen in the X-ray powderdiffraction diagram in FIG. 2. The X-ray powder reflection andintensities (standardised) are shown in Table II.

TABLE II 2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.36 20.24 17 12.177.27 41 12.51 7.07 6 13.13 6.74 7 13.66 6.48 39 14.20 6.23 14 14.60 6.0632 15.03 5.89 5 15.25 5.81 4 15.97 5.54 11 16.51 5.37 13 17.05 5.20 1317.54 5.05 4 17.88 4.96 5 18.65 4.75 22 19.05 4.66 100 19.68 4.51 1120.42 4.35 6 20.84 4.26 4 21.25 4.18 3 21.90 4.06 5 22.42 3.96 92 23.193.83 9 23.70 3.75 16 24.34 3.65 4 24.56 3.62 23 24.89 3.57 16 25.20 3.537 25.36 3.51 7 25.67 3.47 6 26.26 3.39 23 26.59 3.35 12 27.51 3.24 627.71 3.22 6 28.01 3.18 7 28.23 3.16 5 28.57 3.12 3 29.44 3.03 12 30.152.96 4

In Table II above, the value “2-theta [°]” denotes the angle ofdiffraction in degrees and the d-value [A] denotes the specifieddistances in Å between the lattice planes.

The crystalline citrate salt of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanoneis characterised in that the x-ray powder diagram has, inter alia, thecharacteristic values 2-theta=19.1° (100% relative intensity), 22.4°(92% relative intensity), 12.2° (41% relative intensity), 13.7° (39%relative intensity), and 14.6° (32% relative intensity) (which are themost prominent peaks in the diagram of FIG. 2, Table II).

Therefore, according to a first aspect, the invention provides a citratesalt of compound

having the formula

In a second embodiment, salt 1 is in crystalline form.

In a third embodiment, according to any one of the precedingembodiments, the crystalline form of compound 1 shows a X-ray powderdiffraction pattern comprising peaks at the following 2-theta valuesmeasured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40 kV, 40mA: 19.1° and 22.4°.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a X-ray powder diffractionpattern further comprising a peak at 12.2°.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a X-ray powder diffractionpattern further comprising a peak at 13.7°.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a X-ray powder diffractionpattern further comprising a peak at 14.6°.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a X-ray powder diffractionpattern further comprising a peak at 18.7°.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a X-ray powder diffractionpattern further comprising a peak at 24.6°.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a X-ray powder diffractionpattern further comprising a peak at 26.3°.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a Raman spectrum comprisingpeaks at any one or all of the following Raman shifts expressed inwavenumbers in cm⁻¹: 1718, 1242, 731, 662, 553.

In a further embodiment according to any one of the precedingembodiments, the crystalline form shows a melting point of 212±5° C.

The citrate salt 1 may be provided in a pharmaceutical composition.Accordingly, another aspect of the present invention is a pharmaceuticalcomposition containing the salt according to any one of the precedingembodiments optionally together with one or more inert carriers and/ordiluents.

Only one crystalline form has been obtained from several experiments for(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonecitrate.

Hydrobromide Salt of Compound of Formula (I)

The hydrobromide salt of compound of formula (I) is of mediumcrystallinity as demonstrated in the X-ray powder diffraction diagram inFIG. 5. The X-ray powder reflection and intensities (standardised) areshown in Table III.

TABLE III 2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.73 18.67 100 7.6011.62 37 9.48 9.32 15 12.74 6.94 34 14.46 6.12 78 15.25 5.81 62 17.385.10 56 18.16 4.88 17 19.36 4.58 62 20.39 4.35 83 22.01 4.03 17 22.723.91 25 24.05 3.70 37 24.94 3.57 26 25.23 3.53 41 25.65 3.47 27 26.353.38 19 27.25 3.27 19 28.00 3.18 15 28.92 3.08 21 29.49 3.02 15 29.593.02 16

In Table III above, the value “2-theta [°]” denotes the angle ofdiffraction in degrees and the d-value [A] denotes the specifieddistances in A between the lattice planes.

(4-(3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonehydrobromide is characterised in that the x-ray powder diagram has,inter alia, the characteristic values 2-theta=4.7° (100% relativeintensity), 20.4° (83% relative intensity), 14.5° (78% relativeintensity), 15.3° (62% relative intensity), and 19.4° (62% relativeintensity) (which are the most prominent peaks in the diagram of FIG. 5,Table III).

Different polymorphic modifications of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrobromide have beenidentified by X-ray powder diffraction.

Hydrochloride Salt of Compound of Formula (I)

The hydrochloride salt of compound of formula (I) is of mediumcrystallinity as can be seen in the X-ray powder diffraction diagram inFIG. 8. The X-ray powder reflection and intensities (standardised) areshown in Table IV.

TABLE IV 2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.05 21.78 24 4.7418.63 93 7.68 11.50 28 9.49 9.31 19 10.17 8.69 17 12.27 7.21 16 12.856.88 29 13.55 6.53 19 14.05 6.30 22 14.55 6.08 64 15.37 5.76 98 16.095.51 23 16.58 5.34 19 17.52 5.06 100 18.14 4.89 25 19.12 4.64 23 19.534.54 39 20.46 4.34 77 22.16 4.01 23 22.79 3.90 26 23.22 3.83 20 24.133.69 44 25.02 3.56 23 25.42 3.50 24 25.87 3.44 18 26.57 3.35 15 27.393.25 18 28.06 3.18 16 29.07 3.07 18 29.85 3.00 12

In Table IV above, the value “2-theta [° ]” denotes the angle ofdiffraction in degrees and the d-value [A] denotes the specifieddistances in A between the lattice planes.

(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonehydrochloride is characterised in that the x-ray powder diagram has,inter alia, the characteristic values 2-theta =17.5° (100% relativeintensity), 15.4 (98% relative intensity), 4.7° (93% relativeintensity), 20.5° (77% relative intensity), and 14.6° (64% relativeintensity), (which are the most prominent peaks in the diagram of FIG.8, Table IV).

Different polymorphic modifications of(4-((3R,4R)-3-methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanone hydrochloride have beenidentified by X-ray powder diffraction.

Esilate Salt of Compound of Formula (I)

The esilate salt of compound of formula (I) is of high crystallinity ascan be seen in the X-ray powder diffraction diagram in FIG. 11. TheX-ray powder reflection and intensities (standardised) are shown inTable V.

TABLE V 2-theta [°] d-value [Å] Intensity I/I₀ [%] 5.33 16.56 66 7.8711.23 48 9.14 9.67 7 9.97 8.87 24 10.93 8.09 23 12.23 7.23 9 12.43 7.1211 13.26 6.67 83 14.55 6.08 48 14.83 5.97 18 15.07 5.88 10 15.29 5.79 1715.77 5.61 51 16.05 5.52 25 16.18 5.47 19 16.46 5.38 12 16.88 5.25 1017.90 4.95 100 18.32 4.84 34 18.49 4.79 22 19.29 4.60 36 19.44 4.56 4020.03 4.43 63 20.14 4.41 45 20.85 4.26 66 21.08 4.21 11 21.37 4.15 1221.92 4.05 18 22.22 4.00 21 22.49 3.95 16 22.71 3.91 7 23.33 3.81 1023.53 3.78 9 23.79 3.73 8 23.98 3.71 20 24.43 3.64 15 24.68 3.60 1425.00 3.56 17

In Table V above, the value “2-theta [°]” denotes the angle ofdiffraction in degrees and the d-value [Å] denotes the specifieddistances in Å between the lattice planes.

(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanoneesilate is characterised in that the x-ray powder diagram has, interalia, the characteristic values 2-theta=17.9° (100% relative intensity),13.3 (83% relative intensity), 5.3° (66% relative intensity), 20.9° (66%relative intensity), and 20.0° (63% relative intensity) (which are themost prominent peaks in the diagram of FIG. 11, Table V).

Methanesulfonate Salt of Compound of Formula (I)

The methanesulfonate salt of compound of formula (I) is of mediumcrystallinity as can be seen in the X-ray powder diffraction diagram inFIG. 14. The X-ray powder reflection and intensities (standardised) areshown in Table VI.

TABLE VI 2-theta [°] d-value [Å] Intensity I/I₀ [%] 5.36 16.48 30 5.5715.85 27 7.84 11.27 43 9.91 8.92 51 11.05 8.00 21 12.33 7.17 77 13.266.67 26 14.69 6.03 100 14.95 5.92 50 15.78 5.61 20 16.47 5.38 23 17.744.99 53 18.42 4.81 38 19.09 4.65 33 19.29 4.60 41 19.91 4.46 32 20.674.29 55 21.23 4.18 21 22.28 3.99 28 23.74 3.74 16 24.33 3.66 23 24.843.58 15 25.60 3.48 21 29.79 3.00 16 17.74 16.48 30 18.42 15.85 27 19.0911.27 43

In Table VI above, the value “2-theta [°]” denotes the angle ofdiffraction in degrees and the d-value [Å] denotes the specifieddistances in Å between the lattice planes.

(4-((3R,4R)-3-Methoxytetrahydro-pyran-4-ylamino)piperidin-1-yl)(5-methyl-6-(((2R,6S)-6-(p-tolyl)tetrahydro-2H-pyran-2-yl)methylamino)pyrimidin-4-yl)methanonemethanesulfonate is characterised in that the x-ray powder diagram has,inter alia, the characteristic values 2-theta=14.7° (100% relativeintensity), 12.3 (77% relative intensity), 20.7° (55% relativeintensity), 17.7° (53% relative intensity), and 9.9° (51% relativeintensity) (which are the most prominent peaks in the diagram of FIG.14, Table VI).

Thermoanalysis

The thermoanalysis of the crystalline citrate salt 1 shows a meltingpoint=212±5° C. (onset, DSC: 10 K·min⁻¹ heating rate; DSC/TG diagram isshown in FIG. 3). 1.6% weight loss occurs on drying. Consequently, thecitrate salt has a low tendency to absorb solvents (in case of watermeaning low hygroscopicity).

The thermoanalysis of the crystalline hydrobromide salt of compound Ishows a melting point=248±5° C. (onset, DSC: 10 K·min⁻¹ heating rate;DSC/TG diagram is shown in FIG. 6). A broad endothermic effect occursbetween 40-110° C. with concomitant weight loss (2.9% weight loss ondrying).

The thermoanalysis of the crystalline hydrochloride salt of compound Ishows a melting point=233±5° C. (onset, DSC: 10 K·min⁻¹ heating rate;DSC/TG diagram is shown in FIG. 9). A broad endothermic effect occursbetween 40-80° C. A weak endothermic effect occurs between 130-150° C.(2.8% weight loss on drying).

The thermoanalysis of the crystalline esilate salt of compound I shows amelting point=199±5° C. (onset, DSC: 10 K·min⁻¹ heating rate; DSC/TGdiagram is shown in FIG. 12). A weak broad endothermic effect occursbetween 40-100° C. 2.4% loss on drying is correlated with theendothermic effect.

The thermoanalysis of the crystalline methanesulfonate salt of compoundI shows a melting point=226±5° C. (onset, DSC: 10 K·min⁻¹ heating rate;DSC/TG diagram is shown in FIG. 15). A weak broad endothermic effectoccurs between 30-110° C.

Sorption Isotherms

The Sorption isotherm of the crystalline citrate salt 1 shows a wateruptake of 2.6% in the humidity range of 10-80% (diagram shown in FIG.4).

The Sorption isotherm of the crystalline hydrobromide salt of compound Ishows a water uptake of 4.5% in the humidity range of 10-80% (diagramshown in FIG. 7).

The Sorption isotherm of the crystalline hydrochloride salt of compoundI shows a water uptake of 15% in the humidity range of 10-80% (diagramshown in FIG. 10).

The Sorption isotherm of the crystalline esilate salt of compound Ishows a water uptake of 20% in the humidity range of 10-80% (diagramshown in FIG. 13).

The Sorption isotherm of the crystalline methanesulfonate salt ofcompound I shows a water uptake of 30% in the humidity range of 10-80%(diagram shown in FIG. 16).

Summary of Selected Physical Properties for Salts of Compound I

Selected properties of the citrate, hydrobromide, hydrochloride, esilateand methanesulfonate salts of compound I are shown in Table VII.

TABLE VII Salt Form of Compound I Citrate Hydrobromide HydrochlorideEsilate Methanesulfonate Parameter salt Salt Salt Salt Saltcrystallinity high medium medium high Medium melting point 212 ± 5 248 ±5 233 ± 5 199 ± 5 226 ± 5 [° C.] (onset) thermal no broad broad weakbroad weak broad behavior additional endothermic endothermic endothermicendothermic effect effect effect effect effect before 40-110° C. 40-80°C. 40-100° C. 30-110° C. melting weak endothermic effect 130-150° C.loss on 1.6 2.9 2.8 2.4 drying [%] hygroscopic 2.6% 4.5% uptake of 15%uptake 20% uptake 30% uptake behavior uptake of water of water of waterof water (up to 80% r.h.) water deliquescent deliquescent deliquescenthygroscopic 3.4% 20% uptake of 40% uptake 45% uptake 45% uptake behavioruptake of water of water of water of water (up to 90% r.h.) waterdeliquescent deliquescent deliquescent indications no Yes yes no No forpolymorphism

EXAMPLE 2—Biological Activity Data Characterizing Compound I and ItsCitrate Salt 1

Experiments were performed to evaluate the biological activity ofcompound I and its citrate salt 1. A description of the experimentalprocedures and results are provided below.

Part I: Description of Biological Assays Plasma Protein Binding

Dianorm Teflon dialysis cells (micro 0.2) are used. Each cell consistsof a donor (i.e., buffer chamber) and an acceptor chamber (i.e., plasmachamber), separated by an ultrathin semipermeable membrane with a 5 kDamolecular weight cutoff. Stock solutions for each test compound areprepared in DMSO at 1 mM and diluted to a final concentration of 1.0 μM.Aliquots of 200 μl dialysis buffer (100 mM potassium phosphate, pH 7.4)are dispensed into the buffer chamber. Aliquots of 200 μL test compounddialysis solution are dispensed into the plasma chambers. Incubation iscarried out for 2 hours under rotation at 37° C. Then, the dialysate istransferred into reaction tubes. The tubes for the buffer fractioncontain 0.2 mL acetonitril/water (80/20 volume/volume). Aliquots of 25μL of the plasma dialysate are transferred into deep well plates andmixed with 25 μL acetonitril/water (80/20 volume/volume), 25 μL buffer,25 μL calibration solution and 25 μL Internal Standard solution. Proteinprecipitation is done by adding 200 μL acetonitrile. Aliquots of 50 μLof the buffer dialysate are transferred into deep well plates and mixedwith 25 μL blank plasma, 25 μL Internal Standard solution and 200 μLacetonitril. Percent bound is calculated with the formula: %bound=(plasma concentration−buffer concentration/plasmaconcentration)×100.

In Vitro Metabolic Stability

Metabolic degradation of the test compound is assayed in a hepatocytesuspension. Hepatocytes are incubated in an appropriate buffer system.Following a (typically) 30 min preincubation in an incubator (37° C.,10% CO₂) 5 μL of test compound solution (1 μM) are added into 395 μLhepatocyte suspension (cell density in the range 0.25-5 Mio cells/mL,typically 1 Mio cells/mL, final DMSO concentration 0.05%). The cells areincubated for six hours (incubator, orbital shaker) and samples (25 μL)are taken at 0, 0.5, 1, 2, 4 and 6 hours. Samples are transferred intoacetonitrile and pelleted by centrifugation (5 min). The supernatant istransferred to a new 96-deepwell plate, evaporated under nitrogen andresuspended. Decline of parent compound is analyzed by HPLC-MS/MS. CLintis calculated as followsCL_INTRINSIC=Dose/AUC=(CO/CD)/(AUD+clast/k)×1000/60. C0: initialconcentration in the incubation [μM], CD: cell density of vital cells[10e6cells/mL], AUD: area under the data [μM×h], clast: concentration oflast data point [μM], k: slope of the regression line for parent decline[h-1]. The calculated in vitro hepatic intrinsic clearance can be scaledup to the intrinsic in vivo hepatic Clearance and used to predicthepatic in vivo blood clearance (CL) by the use of a liver model (wellstirred model).

-   -   CL_INTRINSIC_INVIVO [ml/min/kg]=(CL_INTRINSIC        [μL/min/10e6cells]×hepatocellularity [10e6 cells/g liver]×liver        factor [g/kg bodyweight])/1000    -   CL [ml/min/kg]=CL_INTRINSIC_INVIVO [ml/min/kg]×hepatic blood        flow [ml/min/kg]/(CL_INTRINSIC_INVIVO [ml/min/kg]+hepatic blood        flow [ml/min/kg])

Pharmacokinetics (Animal Experiments)

The pharmacokinetics of the test compound following single intravenous(IV) or oral (PO) doses were examined in

-   -   female BALB/c mice (average weight: 25 g)    -   male Wistar(Han) rats (average weight: 260 g)    -   male and female Gottingen Minipigs (average weight: 24 kg)    -   male beagle dogs (average weight: 15 kg)        All non-rodent species were fasted overnight prior to dosing,        while mice and rats had food and water available ad libitum. The        p.o. dose of the compound was usually administered as suspension        in 0.5% Natrosol or as a 0.5% Natrosol/0.015% Tween 80        suspension. For i.v. dosing purposes, the doses were applied as        a solution in 0.9% NaCL, or as a solution containing 9.1%        HP-beta Cyclodextrin in water.

Blood was collected by venous sampling and soaking of the blood in EDTAcoated tubes. Samples were collected for up to 48h after administrationof the test compound. Plasma was then separated by centrifugation (5 minby approximately 9000 g at 4° C.). For determination of the testcompound, plasma was transferred into PCR plates. All samples werestored at approximately −20° C. until bioanalytics. The test compoundconcentrations in plasma were determined by HPLC MS/MS. The lower limitof quantification was between 0.5 nmol/L and 1 nmol/L.

hERG-Channel Assay Cells:

HEK (human embryonic kidney) 293 cells were stably transfected with hERGcDNA. Cells determined for use in patch clamp experiments werecultivated without antibiotic.

Pipettes and Solutions

Cells were superfused with a bath solution containing (mM): NaCl (137),KCl (4.0), MgCl₂ (1.0), CaCl₂ (1.8), Glucose (10), HEPES (10), pH 7.4with NaOH. Patch pipettes were made from borosilicate glass tubing(Hilgenberg, Malsfeld, Germany) using a horizontal puller (DMZ-UniversalPuller, Zeitz-Instrumente, Martinsried, Germany) and filled with pipettesolution containing (mM): K-aspartate (130), MgCl₂ (5.0), EGTA (5.0),K₂ATP (4.0), HEPES (10.0), pH 7.2 with KOH. Resistance of themicroelectrodes was in the range between 2 and 5 MΩ.

Stimulation and Recording

Membrane currents were recorded using an EPC-10 patch clamp amplifier(HEKA Electronics, Lambrecht, Germany) and PatchMaster software (HEKA).The current signals were Bessel filtered at 2.5 kHz before beingdigitized at 5 kHz.

hERG-mediated membrane currents were recorded at typically 28° C., usingthe whole-cell configuration of the patch-clamp technique. TransfectedHEK293 cells were clamped at a holding potential of −60 mV andhERG-mediated inactivating tail currents were elicited using a pulsepattern with fixed amplitudes (activation/inactivation: 40 mV for 2000ms; recovery: 120 mV for 2 ms; ramp to 40 mV in 2 ms; inactivating tailcurrent: 40 mV for 50 ms) repeated at 15 s intervals. During eachinter-pulse interval 4 pulses scaled down by a factor of 0.2 wererecorded for a P/n leak subtraction procedure. R_(s) compensation wasemployed up to a level that safely allowed recording devoid of ringing.The remaining uncompensated R_(s) was recorded as well as actualtemperature and holding current.

Compound Preparation and Application

The concentrations of the test item were applied sequentially on each ofthe different cells investigated. A steady state level of baselinecurrent was measured for at least 90 s prior to the application of thefirst test article concentration.

The test item was dissolved in DMSO to yield a stock solution of1000-fold the highest final concentration. This stock was dilutedfurther in DMSO to stock solutions of 1000-fold the remaining finalconcentrations. Final dilutions in extracellular buffer were preparedfreshly from these stocks by a 1:1000 dilution step each before startingthe experiments.

Data Analysis

Peak current amplitudes were measured 3 ms after the ramp to +40 mV. Forbaseline and each concentration the peak currents of the three lastsweeps before application of the next concentration were averaged.Residual currents (I/I₀) were calculated for each cell as the fractionof actual average peak current and average baseline peak current.Current inhibition was expressed as (1−I/I₀)*100%. Current inhibitionfor all cells is reported as mean ±SD. From mean current inhibitiondata, the IC50 is estimated based on the Hill equation using a leastsquares procedure.

In Vitro Phospholipidosis Assay 1. Cell Culture

Cell line: U937. Cell density: 0.5 Mio. cells/mL. Amount of medium: 3mL/well.

2. Materials and Devices

-   Falcon Tissue Culture Flask 175 cm²-   test tubes Sarstedt-   6-well microplates-   laminar flow-   refrigerated centrifuge-   pipettes-   Flow cytometer: Coulter Epics XL/MCL (Beckman Coulter Inc.,    Bullerton, Calif., USA)

3. Medium and Additives 3.1 Preparation of RPMI1640 with 10% FCS and0.005% Gentamicin Media

-   VLE RPMI 1640 medium (1×), store at 2-8° C.

Additives

-   fetal bovine serum, store at −20° C.-   Gentamicin, Gibco® Invitrogen, conc. 10 mg/mL (=1% solution)

Add 56 mL FCS and 2.6 mL Gentamicin to 500 mL RPMI1640. Store theready-to-use medium at 2-8° C.

3.2 Preparation of Formaldehyde Working Solution (conc. 3.7%)

Dilute Formaldehyde 37% in 1×PBS (dilution ratio 1:10) to make a 3.7%working solution, which is stored at 2-8° C.

3.3 Buffer

PBS-Dulbecco (1×) w/o Ca²⁺, Mg²⁺. Store at RT.

4. Dyes for Cell Staining 4.1 Live Cell Staining 4.1.1 Propidium Iodide(PI; Molecular Probes, Eugene, Oreg., USA)

PI stock solution: 1 mg/mL PBS (stored at 4° C. in the dark).

PI ready to use solution: stock solution 1:100 diluted with PBS (freshlyprepared for each experiment).

4.1.2 Nile Red (NR; Molecular Probes, Eugene, Oreg.)

NR stock solution: 1 mg/mL DMSO (stored at 4° C. in the dark).

NR ready to use solution for live cell staining: NR stock solution 1:100diluted with PBS (freshly prepared for each experiment).

4.2 Fixed Cell Staining

Preparation of Nile Red stock solution (conc. 1 mg/mL): solve 1 mg NileRed in 1 mL 100% DMSO, store at 2-8° C.

Preparation of Nile Red working solution for fixed cell staining (conc.1 μg/mL): dilute Nile Red stock solution in 1×PBS (dilution ratio1:1000). The working solution must be prepared and used immediatelybefore staining the cells.

5. Cell Seeding and Treatment

Cell seeding and treatment may be performed as follows:

-   solve the test compounds in 100% DMSO to the 100 fold final    concentration and dilute them according to the experiment planned.-   firstly fill 30 μL of the stock solution in the relevant well of the    6 well plate and re-suspend with-   3 mL cell suspension/well containing 0.5 Mio. cells/mL (final    concentration DMSO=1%).-   use one well per compound and concentration-   incubate 48 hours without changing the medium at 37° C., 5% CO₂ and    95% relative humidity

6. Cell Harvesting

Cell harvesting may be performed as follows:

-   transfer the cell suspension in Sarstedt tubes (on ice)-   centrifugation: 4 min at 130×g, 4° C.; discard the supernatant-   re-suspend in 3 mL PBS per tube (ice cold)-   fill 1 mL of the cell suspension in a Sarstedt tube (on ice) for    flow cytometric determination (0.5 mL for Propidium-iodide and 0.5    mL for Nile Red live cell staining)-   centrifugation of the residual: 4 min. at 130×g, 4° C.; discard the    supernatant-   add 1 mL 3.7% Formaldehyde solution per tube-   fixation for 30 minutes (cells after fixation at RT)-   centrifugation: 4 min at 130×g, RT; discard the supernatant-   re-suspend each tube in 1.3 mL Nile Red working solution for fixed    cell staining-   incubate dye for 5 min-   centrifugation: 4 min at 130×g, RT; discard the supernatant-   re-suspend in 3 mL PBS-   centrifugation: 4 min at 130×g, RT; discard the supernatant-   re-suspend in 0.5 mL PBS (=fraction of Nile Red stained fixed    cells), determination of phospholipidosis using a flow cytometric    method

7. Cell Staining and Flow Cytometric Measurement

3×0.5 mL suspensions of cells are prepared from each sample for flowcytometry measurement (non-fixed cells for viability determination,non-fixed cells and fixed cells for phospholipidosis analysis).

7.1 PI Staining and Flow Cytometric Measurement for ViabilityDetermination

Immediately before measurement, 12.5 μL of the PI ready to use solutionis added per sample (0.5 mL non-fixed cell suspension), which are kepton ice for another 15 min before measurement.

Per sample, ten-thousand (10 000) cells are analyzed at high flow ratefor the following parameter:

-   time to measure 10,000 cells, ungated-   forward scatter (linear) versus sideward scatter (linear), ungated-   yellow fluorescence (λ=568-590 nm; logarithmic) versus cell number    (linear), ungated.

The time to measure 10,000 cells correlates to cell density in thesample.

Cut-off gates for the fluorescence-dependent differentiation betweenlife and dead cells are defined based on the analysis of cell culturemedium plus vehicle exposed Control cells. Cells with a fluorescencelower than the cut-off are defined as viable. Absolute viability of asample is the relation of viable cells to total cell number andexpressed as percentage.

7.2 Nile Red Staining and Flow Cytometric Measurement for PLDetermination 7.2.1 Nile Red Live Cell Staining

Immediately before measurement, 50 μL of the NR ready to use solutionfor live cell staining is added per sample (0.5 mL non-fixed cellsuspension). Samples are kept on ice for another 5 min. Thereafter, theyare washed once with 4 mL PBS (4° C., 250×G for 8 min) and finallyresuspended in 400 μL PBS.

7.2.2 Nile Red Fixed Cell Staining

Description see above (6. Cell harvesting). Both the Nile Red stainednon-fixed cells as well as the Nile Red stained fixed cells are measuredaccording the following procedure.

Per sample, 10,000 cells are analyzed at high flow rate for thefollowing parameter:

-   forward scatter (linear) versus sideward scatter (linear), ungated-   green fluorescence (λ=504-541 nm; logarithmic) versus cell number    (linear), ungated-   far red fluorescence (λ=660-680 nm; logarithmic) versus cell number    (linear), ungated

8. Signal Analysis

Samples of less than 90% relative viability are excluded from analysisof the phospholipidogenic potential of a test compound. Samples with aviability between 90 to 95% are selected for assessment case by casedepending on the consistency of all analyzed parameters and the absolutefluorescence intensity.

For all samples with a viability relative to Control of >90% (based onPI exclusion), the mean absolute fluorescence intensity following NRstaining is calculated for green fluorescence as well as for far redfluorescence.

For each channel, absolute fluorescence intensity of a specific sampleis correlated to the mean absolute fluorescence intensity of all cellculture medium plus vehicle exposed Control cells of the respectiveexperiment. Per channel, relative fluorescence intensity of a sample isthe relation of absolute fluorescence intensity of this sample to themean absolute fluorescence intensity of Controls, which is set at 100,and is expressed as percentage of Control cell fluorescence intensity.

9. Assessment of Phopholipidosis

Assessment of the phospholipidogenic potential of a test compound isdone manually based on the signal intensities at both wavelengths forthe fixed cells as well as for the non-fixed cells.

Part II: Results of Biological Activity Assays for Compound I (FreeBase) and Its Citrate Salt 1

Tables below summarize biological data on compound I and its citratesalt 1, as determined in the assays as described above.

In Vitro Plasma Protein Binding of Compound I

Species Mouse Rat Dog Minipig Human Fraction bound [%] 95.1 68.9 70.460.8 84.7 Fraction unbound [%] 4.9 31.1 29.6 39.2 15.3

In Vitro Metabolic Stability of Compound I in Hepatocyte Incubations

Species Mouse Rat Dog Minipig Human CL intrinsic, in vitro 16.4 8.773.15 2.73 4.11 [μL/min/10e6 cells] CL, in vivo 49 26 14 6.8 7.9[mL/min/kg]

Intravenous Pharmacokinetics of Compound I in Animals

Species Mouse Rat Dog Minipig Animal number/ n = 2f n = 2m n = 3m n =1m/1f gender Intravenous PK parameters (mean values) IV dose (μmol/kg)10 5 5 5 AUC(0-inf) (nM · h) 1990 1490 5990 4310 CL (mL/min/kg) 86.056.1 14.0 20.0 V_(ss) (L/kg) 3.29 5.04 4.94 5.07 MRT_(disp) (h) 0.6231.49 6.40 4.15

Oral Pharmacokinetics of Compound I in Animals

Species Mouse Rat Dog Minipig Animal number/ n = 3m/0f n = 3m/0f n =3m/0f n = 3m/0f gender Oral PK parameters (mean values) Oral Dose(μmol/kg) 20 20 5 not done C_(max) (nM) 974 580 317 not done t_(max) (h)1.00 1.50 0.917 not done AUC(0-inf) (nM · h) 3160 2270 1500 not doneMRT_(tot) (h) 3.99 5.49 5.77 not done F (%) 79 38 25 Not calculated

Oral Pharmacokinetics of Citrate Salt 1 in Rats

Species Rat Animal number/ n = 3m/0f gender Oral PK parameters ^(c)(mean values) Oral Dose (μmol/kg) 20 C_(max) (nM) 454 t_(max) (h) 1.08AUC(0-inf) (nM · h) 1710 MRT_(tot) (h) 3.3

Inhibition of hERG-Mediated Potassium Current

Compound I inhibited the hERG-mediated potassium current with IC₅₀>30 μM(12% inhibition at 10 μM, 28% inhibition at 30 μM).

In vitro Phospholipidosis Assay

Compound I shows the propensitiy to be phospholipidogenic in the invitro Phospholipidosis assay; the lowest phospholipidogenicconcentration of compound I in this in vitro assay is 200 μM.

INCORPORATION BY REFERENCE

The entire disclosure of each of the patent documents and scientificarticles referred to herein is incorporated by reference for allpurposes.

EQUIVALENTS

The invention may be embodied in other specific forms without departingfrom the spirit or essential characteristics thereof The foregoingembodiments are therefore to be considered in all respects illustrativerather than limiting the invention described herein. Scope of theinvention is thus indicated by the appended claims rather than by theforegoing description, and all changes that come within the meaning andrange of equivalency of the claims are intended to be embraced therein.

What is claimed is:
 1. A method of inhibiting CCR2 activity in a patientsuffering from pain, comprising administering to the patient in needthereof a therapeutically effective amount of a compound of Formula 1:


2. The method of claim 1, wherein the pain is inflammatory pain.
 3. Themethod of claim 1, wherein the pain is chronic pain.
 4. The method ofclaim 1, wherein the pain is due to osteoarthritis.
 5. The method ofclaim 1, wherein the compound of Formula 1 is in crystalline form. 6.The method of claim 2, wherein the compound of Formula 1 is incrystalline form.
 7. The method of claim 3, wherein the compound ofFormula 1 is in crystalline form.
 8. The method of claim 4, wherein thecompound of Formula 1 is in crystalline form.
 9. The method of claim 5,wherein the crystalline form is characterized by a X-ray powderdiffraction pattern comprising peaks at the following 2-theta valuesmeasured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40 kV, 40mA: 19.1° and 22.4°.
 10. The method of claim 6, wherein the crystallineform is characterized by a X-ray powder diffraction pattern comprisingpeaks at the following 2-theta values measured using monochromatic CuKα1radiation of λ=1.54056 Å, 40 kV, 40 mA: 19.1° and 22.4°.
 11. The methodof claim 7, wherein the crystalline form is characterized by a X-raypowder diffraction pattern comprising peaks at the following 2-thetavalues measured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40kV, 40 mA: 19.1° and 22.4°.
 12. The method of claim 8, wherein thecrystalline form is characterized by a X-ray powder diffraction patterncomprising peaks at the following 2-theta values measured usingmonochromatic CuKα1 radiation of λ=1.54056 Å, 40 kV, 40 mA: 19.1° and22.4°.
 13. The method of claim 5, wherein the crystalline form exhibitsa X-ray powder diffraction pattern comprising peaks at the following2-theta values measured using monochromatic CuKα1 radiation of λ=1.54056Å, 40kV, 40 mA: 12.2±0.2, 13.7±0.2, 14.6±0.2, 19.1±0.2, and 22.4±0.2.14. The method of claim 6, wherein the crystalline form exhibits a X-raypowder diffraction pattern comprising peaks at the following 2-thetavalues measured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40kV, 40 mA: 12.2±0.2, 13.7±0.2, 14.6±0.2, 19.1±0.2, and 22.4±0.2.
 15. Themethod of claim 7, wherein the crystalline form exhibits a X-ray powderdiffraction pattern comprising peaks at the following 2-theta valuesmeasured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40 kV, 40mA: 12.2±0.2, 13.7±0.2, 14.6±0.2, 19.1±0.2, and 22.4±0.2.
 16. The methodof claim 8 wherein the crystalline form exhibits a X-ray powderdiffraction pattern comprising peaks at the following 2-theta valuesmeasured using monochromatic CuKα1 radiation of λ=1.54056 Å, 40 kV, 40mA: 12.2±0.2, 13.7±0.2, 14.6±0.2, 19.1±0.2, and 22.4±0.2.
 17. The methodof claim 5, wherein the crystalline form is characterized by thefollowing X-ray powder diffraction pattern expressed in terms ofdiffraction angle 2θ, inter-planar distances d, and relative intensity(expressed as a percentage with respect to the most intense peak):2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.36 20.24 17 12.17 7.27 4112.51 7.07 6 13.13 6.74 7 13.66 6.48 39 14.20 6.23 14 14.60 6.06 3215.03 5.89 5 15.25 5.81 4 15.97 5.54 11 16.51 5.37 13 17.05 5.20 1317.54 5.05 4 17.88 4.96 5 18.65 4.75 22 19.05 4.66 100 19.68 4.51 1120.42 4.35 6 20.84 4.26 4 21.25 4.18 3 21.90 4.06 5 22.42 3.96 92 23.193.83 9 23.70 3.75 16 24.34 3.65 4 24.56 3.62 23 24.89 3.57 16 25.20 3.537 25.36 3.51 7 25.67 3.47 6 26.26 3.39 23 26.59 3.35 12 27.51 3.24 627.71 3.22 6 28.01 3.18 7 28.23 3.16 5 28.57 3.12 3 29.44 3.03 12 30.152.96 4


18. The method of claim 6, wherein the crystalline form is characterizedby the following X-ray powder diffraction pattern expressed in terms ofdiffraction angle 2θ, inter-planar distances d, and relative intensity(expressed as a percentage with respect to the most intense peak):2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.36 20.24 17 12.17 7.27 4112.51 7.07 6 13.13 6.74 7 13.66 6.48 39 14.20 6.23 14 14.60 6.06 3215.03 5.89 5 15.25 5.81 4 15.97 5.54 11 16.51 5.37 13 17.05 5.20 1317.54 5.05 4 17.88 4.96 5 18.65 4.75 22 19.05 4.66 100 19.68 4.51 1120.42 4.35 6 20.84 4.26 4 21.25 4.18 3 21.90 4.06 5 22.42 3.96 92 23.193.83 9 23.70 3.75 16 24.34 3.65 4 24.56 3.62 23 24.89 3.57 16 25.20 3.537 25.36 3.51 7 25.67 3.47 6 26.26 3.39 23 26.59 3.35 12 27.51 3.24 627.71 3.22 6 28.01 3.18 7 28.23 3.16 5 28.57 3.12 3 29.44 3.03 12 30.152.96 4


19. The method of claim 7, wherein the crystalline form is characterizedby the following X-ray powder diffraction pattern expressed in terms ofdiffraction angle 2θ, inter-planar distances d, and relative intensity(expressed as a percentage with respect to the most intense peak):2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.36 20.24 17 12.17 7.27 4112.51 7.07 6 13.13 6.74 7 13.66 6.48 39 14.20 6.23 14 14.60 6.06 3215.03 5.89 5 15.25 5.81 4 15.97 5.54 11 16.51 5.37 13 17.05 5.20 1317.54 5.05 4 17.88 4.96 5 18.65 4.75 22 19.05 4.66 100 19.68 4.51 1120.42 4.35 6 20.84 4.26 4 21.25 4.18 3 21.90 4.06 5 22.42 3.96 92 23.193.83 9 23.70 3.75 16 24.34 3.65 4 24.56 3.62 23 24.89 3.57 16 25.20 3.537 25.36 3.51 7 25.67 3.47 6 26.26 3.39 23 26.59 3.35 12 27.51 3.24 627.71 3.22 6 28.01 3.18 7 28.23 3.16 5 28.57 3.12 3 29.44 3.03 12 30.152.96 4


20. The method of claim 8, wherein the crystalline form is characterizedby the following X-ray powder diffraction pattern expressed in terms ofdiffraction angle 2θ, inter-planar distances d, and relative intensity(expressed as a percentage with respect to the most intense peak):2-theta [°] d-value [Å] Intensity I/I₀ [%] 4.36 20.24 17 12.17 7.27 4112.51 7.07 6 13.13 6.74 7 13.66 6.48 39 14.20 6.23 14 14.60 6.06 3215.03 5.89 5 15.25 5.81 4 15.97 5.54 11 16.51 5.37 13 17.05 5.20 1317.54 5.05 4 17.88 4.96 5 18.65 4.75 22 19.05 4.66 100 19.68 4.51 1120.42 4.35 6 20.84 4.26 4 21.25 4.18 3 21.90 4.06 5 22.42 3.96 92 23.193.83 9 23.70 3.75 16 24.34 3.65 4 24.56 3.62 23 24.89 3.57 16 25.20 3.537 25.36 3.51 7 25.67 3.47 6 26.26 3.39 23 26.59 3.35 12 27.51 3.24 627.71 3.22 6 28.01 3.18 7 28.23 3.16 5 28.57 3.12 3 29.44 3.03 12 30.152.96 4